Enzyme/immunofluorescent assay for anti-treponemal antibodies

Abstract

A method for the determination of anti-treponemal antibody in a test sample comprises contacting a substrate for the anti-treponemal antibody with sample; treating the contacted substrate with labeled antihuman Ig antibody selected from (a) a mixture comprising enzyme labeled antihuman Ig antibody and fluorescent labeled antihuman Ig antibody, (b) antihuman Ig antibody labeled with an enzyme and a fluorescent label, and (c) fluorescent labeled antihuman Ig antibody to which enzyme labeled antibody against the animal species from which the antibody used in the fluorescent labeled antibody was derived is added subsequently, and (d) enzyme labeled antihuman Ig antibody to which fluorescent labeled antibody against the animal species from which the antibody used in the enzyme labeled antibody was derived is subsequently added; determining the enzyme activity of the treated substrate; and determining the immunofluorescent patterns in substrates exhibiting enzyme activity. The method is useful for the rapid screening of anti-treponemal antibodies for the diagnosis of syphilis.

What is claimed is: 
     
       1. A method for the determination of anti-treponemal antibody in a test sample, comprising: providing a substrate for the anti-treponemal antibody; contacting said substrate with a test sample; treating said contacted substrate with labeled antihuman immunoglobulin antibody, said labeled antihuman immunoglobulin antibody selected from the group consisting of: (a) a mixture comprising enzyme labeled antihuman immunoglobulin antibody and fluorescent labeled antihuman immunoglobulin antibody;   (b) antihuman immunoglobulin antibody labeled with an enzyme and a fluorescent label;   (c) fluorescent labeled antihuman immunoglobulin antibody to which enzyme labeled antibody against the animal species from which the antibody used in the fluorescent labeled antibody was derived is subsequently added; and   (d) enzyme labeled antihuman immunoglobulin antibody to which fluorescent labeled antibody against the animal species from which the antibody used in the fluorescent labeled antibody was derived is subsequently added;     analyzing of the treated substrate to determine if it has enzyme activity; and   determining the immunofluorescent pattern in substrates exhibiting enzyme activity.   
     
     
       2. The method according to claim 1 wherein said labeled antihuman antibody comprises (a). 
     
     
       3. The method according to claim 1 wherein said labeled antihuman antibody comprises (b). 
     
     
       4. The method according to claim 1 wherein said labeled antihuman antibody comprises (c). 
     
     
       5. The method according to claim 1 wherein said labeled antihuman antibody comprises (d). 
     
     
       6. The method according to claim 2 wherein said enzyme is horseradish peroxidase. 
     
     
       7. The method according to claim 3 wherein said enzyme is horseradish peroxidase. 
     
     
       8. The method according to claim 3 wherein said enzyme is horseradish peroxidase. 
     
     
       9. The method according to claim 2 wherein said fluorescent label is fluorescein. 
     
     
       10. The method according to claim 3 wherein said fluorescent label is fluorescein. 
     
     
       11. The method according to claim 4 wherein said fluorescent label is fluorescein. 
     
     
       12. The method according to claim 2 wherein said enzyme is horseradish peroxidase and said fluorescent label is fluorescein. 
     
     
       13. The method according to claim 3 wherein said enzyme is horseradish peroxidase and said fluorescent label is fluorescein. 
     
     
       14. The method according to claim 4 wherein said enzyme is horseradish peroxidase and said fluorescent label is fluorescein. 
     
     
       15. The method according to claim 1 wherein said antihuman antibody is antihuman IgG. 
     
     
       16. The method according to claim 1 wherein said substrate comprises Treponema pallidum cells.